Comprehensive mutagenesis maps the effect of all single-codon mutations in the AAV2 gene on AAV production.

Recombinant adeno-associated viruses (rAAVs) are the predominant gene therapy vector. Several rAAV vectored therapies have achieved regulatory approval, but production of sufficient rAAV quantities remains difficult. The AAV Rep proteins, which are essential for genome replication and packaging, represent a promising engineering target for improvement of rAAV production but remain underexplored.

Deep diversification of an AAV capsid protein by machine learning.

Modern experimental technologies can assay large numbers of biological sequences, but engineered protein libraries rarely exceed the sequence diversity of natural protein families. Machine learning (ML) models trained directly on experimental data without biophysical modeling provide one route to accessing the full potential diversity of engineered proteins. Here we apply deep learning to design highly diverse adeno-associated virus 2 (AAV2) capsid protein variants that remain viable for packaging of a DNA payload.

Comprehensive AAV capsid fitness landscape reveals a viral gene and enables machine-guided design.

Adeno-associated virus (AAV) capsids can deliver transformative gene therapies, but our understanding of AAV biology remains incomplete. We generated the complete first-order AAV2 capsid fitness landscape, characterizing all single-codon substitutions, insertions, and deletions across multiple functions relevant for in vivo delivery. We discovered a frameshifted gene in the VP1 region that expresses a membrane-associated accessory protein that limits AAV production through competitive exclusion.

Reversed phase HPLC with high temperature ethanol/water mobile phases as a green alternative method for the estimation of octanol/water partition coefficients.

High temperature ethanol/water was explored as a green eluent in the reversed-phase liquid chromatographic approximation of pure water retention (log k) and subsequent estimation of the octanol/water partition coefficient (log P) via the Collander equation and the Leave-One-Out method. As part of this work, linear solvation energy relationships were employed to compare the log k extrapolated systems based on high temperature ethanol/water, ambient acetonitrile/water, and ambient methanol/water mobile phases. Based on the comparisons of the three organic modifiers, high temperature ethanol/water mobile phases were observed to provide the best estimation of log P.

Identifying the educational needs of physicians in pediatric epilepsy in order to improve care: results from a needs assessment in Germany, Spain, and the United States.

The objective of this study was to gather evidence-based data on the educational needs of neuropediatricians. A needs assessment was conducted to identify the clinical challenges of physicians when diagnosing, medically treating, and managing pediatric patients with epilepsy; which could be addressed through educational interventions. A two-phase mixed-methods approach was used to conduct the needs assessment in Germany, Spain, and the US.

A Liquid Chromatography/Mass Spectrometry Method for Screening Disulfide Tethering Fragments.

We report the refinement of a high-throughput, liquid chromatography/mass spectrometry (LC/MS)-based screening method for the identification of covalent small-molecule binders to proteins. Using a custom library of 1600 disulfide-capped fragments targeting surface cysteine residues, we optimize sample preparation, chromatography, and ionization conditions to maximize the reliability and flexibility of the approach. Data collection at a rate of 84 s per sample balances speed with reliability for sustained screening over multiple, diverse projects run over a 24-month period.

Characterization of the polarity of subcritical water.

The polarity of subcritical water was studied solvatochromically with betaine dye (33) across a temperature range of 30°C-180°C and a pressure range of 13.8bar (200psi) to 124bar (1800psi). It was observed that temperature has a greater effect than pressure on the polarity of subcritical water.

In situ repair of vagus nerve stimulator lead damage: technical note.

Vagus nerve stimulators (VNSs) are currently an accepted treatment for intractable epilepsy not amenable to ablative surgery. Battery death and lead damage are the main reasons for reoperation in patients with VNSs. In general, any damage to the lead requires revision surgery to remove the helical electrodes from the vagus nerve and replace the electrode array and wire.

An alternative practice model: residents transform continuity clinic and become systems thinkers.

Background: A changing health care environment has created a need for physicians trained in health system improvement. Residency programs have struggled to teach and assess practice-based learning and improvement and systems-based practice competencies, particularly within ambulatory settings.

Intervention: We describe a resident-created and resident-led quality and practice-improvement council in an internal medicine continuity clinic.

Nurses' perceptions of simulation-based interprofessional training program for rapid response and code blue events.

Following completion of an interprofessional simulation program for rapid response and code blue events, we explored hospital unit nurses' perspectives of the training, through a mixed-methods analysis. The results of this study advocate for the use of simulation training in preparing nurses and promoting communication among team members, effective teamwork, and early recognition of clinically deteriorating patients. This study provides support for the implementation and continued use of simulation interprofessional programs in hospital settings.

Systems-based practice in graduate medical education: systems thinking as the missing foundational construct.

Background: Since 2001, residencies have struggled with teaching and assessing systems-based practice (SBP). One major obstacle may be that the competency alone is not sufficient to support assessment. We believe the foundational construct underlying SBP is systems thinking, absent from the current Accreditation Council for Graduate Medical Education competency language.

Retention mechanism of a cholesterol-coated C18 stationary phase: van't Hoff and Linear Solvation Energy Relationships (LSER) approaches.

This study examines the effect of temperature on the dynamic cholesterol coating of a C18 stationary phase and the effect of this coating on the retention mechanism. In general, an increase in temperature results in a decrease in the mass of cholesterol coated on the stationary phase. Typically, an increase in temperature from 25°C to 55°C results in a nearly 60% reduction in the mass of cholesterol loaded.

What can medical education learn from the neurobiology of learning?

The last several decades have seen a large increase in knowledge of the underlying biological mechanisms that serve learning and memory. The insights gleaned from neurobiological and cognitive neuroscientific experimentation in humans and in animal models have identified many of the processes at the molecular, cellular, and systems levels that occur during learning and the formation, storage, and recall of memories. Moreover, with the advent of noninvasive technologies to monitor patterns of neural activity during various forms of human cognition, the efficacy of different strategies for effective teaching can be compared.

Enhancing competency in professionalism: targeting resident advance directive education.

Background: Education about advance directives typically is incorporated into medical school curricula and is not commonly offered in residency. Residents' experiences with advance directives are generally random, nonstandardized, and difficult to assess. In 2008, an advance directive curriculum was developed by the Scott & White/Texas A&M University System Health Science Center College of Medicine (S&W/Texas A&M) internal medicine residency program and the hospital's legal department.

Students learn systems-based care and facilitate system change as stakeholders in a free clinic experience.

Systems-based practice (SBP) is rarely taught or evaluated during medical school, yet is one of the required competencies once students enter residency. We believe Texas A&M College of Medicine students learn about systems issues informally, as they care for patients at a free clinic in Temple, TX. The mandatory free clinic rotation is part of the Internal Medicine clerkship and does not include formal instruction in SBP.

The patient panel conference experience: what patients can teach our residents about competency issues.

Purpose: In 2007, the Scott & White/Texas A&M HSC College of Medicine began requiring all internal medicine residents to attend quarterly patient panel conferences, during which former Scott & White patients speak frankly about their inpatient and outpatient experiences. The main purpose of this mixed-methods pilot study was to determine whether residents' competency education could be enhanced via the conferences.

Method: Of the 54 internal medicine residents in the residency program, 31, 39, and 41 participated in three patient panel conferences, respectively, between December 2007 and August 2008.

Stability and selectivity of a cholesterol-coated C18 stationary phase.

This paper details the use of cholesterol as a mobile phase additive and stationary phase complexing agent in reversed-phase liquid chromatography. Cholesterol loading onto a typical C18 stationary phase is examined. It is found that, when using a standard 150 mm x 4.

Using simulation to orient code blue teams to a new hospital facility.

Objectives: Prompt and successful cardiopulmonary resuscitation during a sudden cardiac arrest can be hindered by multiple variables, ie, ineffective communication, stress, lack of training, and an unfamiliar environment, such as a new hospital facility. The main objective of the study was to use high-fidelity simulations to orient Code Blue Teams (CBTs) to critical events in a new hospital facility. A secondary objective was to elucidate factors that may have contributed to responses by debriefing teams.

Procedural and interpretive skills of medical students: experiences and attitudes of fourth-year students.

Background: Recent data do not exist regarding fourth-year medical students' performance of and attitudes toward procedural and interpretive skills, and how these differ from third-year students'.

Method: Cross-sectional survey conducted in February 2006 of 122 fourth-year students from seven U.S.

The effect of two different faculty development interventions on third-year clerkship performance evaluations.

Background And Objectives: Clinical performance evaluations of medical students often fail to identify significant deficiencies. Many physicians are unwilling to give a poor or failing performance evaluation. Consequently, many clinical rotation grades are inflated and do not reflect actual student performance.

Complying with ACGME resident duty hours restrictions: restructuring the 80-hour workweek to enhance education and patient safety at Texas A&M/Scott & White Memorial Hospital.

Compliance with the Accreditation Council for Graduate Medical Education resident duty hours rules has created unique educational and patient-care challenges for the general medicine inpatient teaching (GMIT) teams at Texas A&M/Scott & White Memorial Hospital, including multiple patient hand-offs, multiple resident absences during teaching time, and loss of continuity of care for individual patients, all of which may have compromised patient safety. The Texas A&M/Scott & White Memorial Hospital internal medicine residency program initially complied with the duty hours rules by having residents take call every fourth night, followed by a six-hour post-call day. This system proved to be inefficient because it significantly disrupted patient care and resident education.